Quality and safety of bovine clones and their products.

A multidisciplinary research programme was developed to get a scientific expertise for the quality assessment of products obtained from cloned livestock. Thirty-seven bovine Holstein female clones of five different genotypes and their products were analysed in comparison with 38 control animals obtained by conventional artificial insemination and raised under the same conditions at the same experimental farm. Animal evaluation included over 150 criteria and more than 10 000 measurements to check the physiological status and health over a 3-year period. All the parameters studied were in the normal range for age and breed, but some significant differences were detected between clone and control groups in terms of delayed onset of puberty in clones, higher neutrophil counts in haematology or lower biochemical plasma concentrations of gamma glutamyl transferase. Milk and meat analyses were conformable to expected values. We, however, found some differences in fatty acid (FA) composition of milk and muscle suggesting a possible deviation in lipid metabolism as assessed by higher delta-9 desaturase activity indexes in both milk and muscles from clones compared with controls. Repeated muscle biopsies in the semitendinosus muscle of the same animals demonstrated a higher oxidative activity in muscle of young clones (8 months of age) compared with controls, suggesting a delayed muscle maturation in clones. Nutritional evaluation of milk and meat using the rat feeding trials did not show any difference between clone and control products for food intake, growth rate, body composition of the rats, nor for possible allergenicity. Possible reactivation of bovine endogenous retroviruses (BERVs) was analysed and compared between normal and cloned cattle. As expected, these BERV sequences are not transcribed and no RNA was detected in the blood of clones, donor animals or controls; therefore, it may be assumed that the sanitary risk associated with BERV sequences is not higher in cattle derived from somatic nuclear transfer than in cattle born from conventional reproduction. Our results confirm that the quality and safety of products (milk and meat) from adult and clinically healthy cloned cattle is globally similar to normal animals. However, from a strictly biological point of view, the slightly delayed maturation we observed in the muscle of clones together with some marginal differences identified in FA composition of both muscle and milk, point to the need for more refined analysis to totally exclude any risks from the consumption of those products.

Health status of cloned cattle at different ages.

The procedure of somatic cloning is associated with important losses during pregnancy and in the perinatal period, reducing the overall efficacy to less than 5% in most cases. A mean of 30% of the cloned calves die before reaching 6 months of age with a wide range of pathologies, including, for the most common, respiratory failure, abnormal kidney development, liver steatosis. Heart and liver weight in relation to body weight are also increased. Surviving animals, although mostly clinically normal, differ from controls obtained by artificial insemination (AI) within the first 1-2 months, to become undistinguishable from them thereafter. Hemoglobin concentrations, for instance, are lower, and leptin concentrations are elevated. In response to the lack of prospective studies addressing the health of adult clones, a long-term, 3-4-year study is currently being conducted to assess the health of mature bovine clones at INRA. Preliminary results over 1 year of study do not show any statistical difference between groups for hematological parameters.

Estrus synchronization in beef cows: comparison between GnRH+PGF2alpha+GnRH and PRID+PGF2alpha+eCG.

The aim of this study was to compare two protocols for estrus synchronization in suckled beef cows over a 2 years period. The population studied consisted of 172 Charolais and 168 Limousin cows from 12 and 14 beef herds, respectively. In each herd, cows were allotted to groups according to parity, body condition score and calving difficulty. Cows in Group 1 (n=174) received PRID on Day-8 with estradiol benzoate (10mg, vaginal capsule), dinoprost on Day-4 (25mg i.m.), eCG on Day 2 (500 IU i.m.). The PRID was removed on Day-2 and cows were inseminated on Day 0, 56 h after PRID was removed. Cows in Group 2 (n=166) received GnRH on Day-10 (100 microg i.m.), dinoprost on Day-3 (25mg i.m.) and GnRH on Day-1 (100 microg i.m.), and were inseminated on Day 0, 16-24h after the last GnRH treatment. Plasma progesterone concentrations were measured to determine cyclicity prior to treatment (Days-20 and -10), to confirm the occurrence of ovulation (Days 0 and 10) and to determine the apparent early pregnancy rate (Days 0, 10 and 24). Pregnancy diagnosis was performed by ultrasonography between Days 35 and 45. The effects of various factors on ovulation, apparent early pregnancy and pregnancy rates were studied using logistic mixed models. There was no significant difference between Groups 1 and 2, respectively, for the cyclicity rate before treatment (80.5% versus 80.1%), for apparent pregnancy rate on Day 24 (62.1% versus 54.8%, P=0.09) and for pregnancy rate on Days 35-45 (53.8% versus 46.3%, P=0.16). Ovulation rate was higher (P<0.01) in Group 1 (90.8%) than in Group 2 (77.1%) and was affected by cyclicity prior to treatment in Group 2 but not in Group 1 (Group 1: 88.2% in anestrous cows versus 91.4% in cyclic cows; Group 2: 45.5% in anestrous cows versus 85.0% in cyclic cows, P interaction=0.05). Apparent pregnancy rates on Day 24 were influenced by the year of study (52.4% versus 68.8%, OR=2.12, P<0.01) and by the cyclicity before treatment (anestrous cows 46.3% versus cyclic cows 61.5%, OR=1.86, P<0.05). Pregnancy rates at 35-45 days were influenced by the year of study (44.2% versus 59.8%, OR=1.92, P<0.01). In conclusion, although pregnancy rates were similar for the two treatments, the combination of GnRH+PGF2alpha+GnRH in suckled beef cows induced a lower rate of ovulation than treatment with PRID+PGF2alpha, particularly in anestrous cows.

Cyclooxygenase-2 is expressed by cumulus cells during oocyte maturation in cattle.

Prostaglandins could be involved in various aspects of final differentiation of ovarian follicles. Prostaglandins are generated by the cyclooxygenase (cox) pathway. Until now, the expression pattern of isoforms cox-1 and cox-2 of cyclooxygenase in bovine cumulus-oocyte complexes (COCs) was unknown. Using immunodetection procedure, we demonstrated in the present study that cox-2 was expressed by cumulus cells during in vivo and in vitro maturation. Time course induction of cox-2 expression was investigated during in vitro maturation using Western blot analysis. Specific signal of cox-2 was markedly evidenced from 6 hr of culture and increased to reach a maximal level at 24 hr of culture. In vitro, cox-2 expression in COCs was associated with increased concentrations of PGE(2) and PGF(2alpha) in the maturation medium. In addition, the effects of culture conditions on cox-2 expression was considered using RT-PCR and Western-blot analysis. We demonstrated that the addition of 10 ng/ml of EGF to TCM199 clearly increased the expression level of cox-2 mRNA and protein. Higher levels of in vitro cox-2 expression was associated with greater rates of cumulus expansion and oocytes at metaphase II at 24 hr of culture. In conclusion, our present results suggest that cox-2 expression in cumulus cells may be involved in differentiation of COCs that occurs during oocyte maturation.

Influence of the type of energy supply on lh secretion, follicular growth and response to estrus synchronization treatment in feed-restricted suckler beef cows.

The present experiment aimed to compare the efficiency of supplementation (+17.5 MJ Net Energy/d starting 47 +/- 4 days after calving) with concentrate (CS, maize grain, n = 10) or with forage (FS, maize silage, n = 10) in estrus-synchronized (Norgestomet implant 10 days inserted 60 +/- 4 days postpartum + PMSG at implant removal) beef cows previously restricted (47 MJ Net Energy/d, 785 g CP/d, 70% of requirements). The type of diet had no significant effect on basal LH concentrations (CS: 0.18 +/- 0.12 vs FS: 0.11+/- 0.02 ng/mL), LH pulse frequency (CS : 0.7 +/- 0.3 vs FS: 0.8 +/- 0.2 pulse/10 h), LH pulse amplitude (CS: 0.55 +/- 0.50 vs FS : 0.62 +/- 0.50 ng/mL) or estradiol (E2) concentrations (CS: 3.3 +/- 0.8 vs FS: 4.6+ /- 0.8 pg/mL) 13 days after the beginning of energy supplementation. No differences between CS and FS cows were observed for the number of small, medium and large follicles nor on the size of the largest follicle from 11 days before implant insertion to implant removal (IR). After IR, an LH surge was observed in 2 of the CS and 4 of the FS cows. The type of energy supplementation had no significant effect on LH (CS: 0.16 +/- 0.06 ng/mL vs FS 0.48 +/- 0.06 ng/mL; P > 0.05) or on estradiol concentrations (CS : 7.8 +/- 0.2 vs FS : 8.9 +/- 0.2 pg/mL, P > 0.10) measured hourly from 29 to 49 h after IR. Cows that ovulated after IR tended to have higher E2 concentrations than cows that did not ovulate (9.4 +/- 0.2 vs 6.3 +/- 0.2 pg/mL, P = 0.08). Similar ovulation and pregnancy rates were observed in CS and FS cows (CS: 6/10 vs FS: 7/10 and CS: 6/10 vs FS: 5/10 respectively, P > 0.05). To conclude, energy supplementation with forage was as effective as energy supplementation with concentrate to influence follicular growth, ovulation and pregnancy percentage after estrus synchronization treatment in diet-restricted beef cows.

Estimation of Neospora caninum seroprevalence in dairy cattle from Normandy, France.

An epidemiological study was conducted in Orne (France) on randomly selected dairy herds (42 herds including 1,924 cows and heifers, which were at least 15 months old). The aim was primarily to estimate the seroprevalence of Neospora caninum infection from two blood samples per cow, using an enzyme-linked immunosorbent assay (ELISA) for N. caninum (one positive result indicating infection). The second aim was to test the association between some individual and herd factors and N. caninum seropositivity with a logistic model including a random term effect. The prevalence was estimated at 5.6% (107 seropositive animals). At least 27 of the 42 herds had one seropositive cow or heifer. The intra-herd seroprevalence varied from 1.1 to 8% for 18 positive herds (66.7%). Dogs were present in 36 farms and 104 of the 107 seropositive animals were exposed to them. The factors associated with individual seropositivity were the presence of cats (OR = 0.17; P < 0.001), dogs (OR = 4.35; P = 0.02), rabbits and/or ducks (OR = 2.10; P = 0.04), long calving periods (12 months) (OR = 0.44; P = 0.007), tethered housing (OR = 2.50; P = 0.01), somatic cell counts (200-400 x 10(3) cells/mL) (OR = 0.24; P < 0.001) and pond water supply (OR = 2.43; P = 0.04). In conclusion, the animal and intra-herd seroprevalences were low in dairy cows from Normandy, France.

Postpartum subestrus in dairy cows: comparison of treatment with prostaglandin F2 alpha or GnRH + prostaglandin F2 alpha + GnRH.

Two experiments (Experiment 1, 185 cows in 1996/97; Experiment 2, 168 cows in 1997/98) were conducted with Prim Holstein dairy cattle in the Mayenne region of France to investigate subestrus. Cows which had not been observed in estrus since calving were allocated alternately to treatment groups between 60 and 90 d post partum as follows: Experiment 1-Group 1: GnRH (Day 0, 100 micrograms i.m.), PGF2 alpha (Day 7, 25 mg i.m.), GnRH (Day 9, 100 micrograms i.m.) and AI (Day 10); Group 2: PGF2 alpha (Day 0, 25 mg i.m.), AI at estrus, or, if estrus was not observed, a second PGF2 alpha injection on Day 13, and AI on Day 16 and Day 17. Treatments in Experiment 2 were as follows: Group 1: as Experiment 1-Group 1 but AI at the observed estrus after Day 0, or at Day 10 if estrus was not observed; Group 2: as Experiment 1--Group 2, however, if a second PGF2 alpha injection was given on Day 13, AI at the observed estrus. Progesterone was measured in serum at Day 0 and in milk at AI. Pregnancy diagnosis was performed by measuring bovine pregnancy-specific protein B (bPSPB; Day 50 +/- 3) and confirmed by ultrasonography when the result was doubtful. In Experiment 1, farmers observed 47/101 (46.9%) Group 1 cows in estrus, 33/91 cows on Day 10 and 10 cows before Day 10. The progesterone concentrations were compatible with estrus in 69/86 (80%) cows on Day 10. In Group 2, 36/83 (43.4%) cows were inseminated after the first PGF2 alpha injection. After the second PGF2 alpha injection, only 29/43 (67%) cows had a low progesterone concentration at AI. Pregnancy rates were 36.1 and 32.5% for Groups 1 and 2, respectively. In Experiment 2, estrus was observed in 31/93 (33.7%) Group 1 cows. In Group 2, 51/75 (66%) cows were inseminated after the first injection of PGF2 alpha, 13/75 (17.3%) cows after the second injection, while 11/75 (14.7%) were not observed in estrus. Pregnancy rates were 53.7 and 53.3% in Groups 1 and 2, respectively. In conclusion, it is recommended that subestrus be treated with PGF2 alpha followed by AI at the observed estrus when estrus detection is good, while the use of GnRH + PGF2 alpha + GnRH is recommended when estrus detection is poor.

The fertility of autumn calving suckler beef cows is increased by the addition of prostaglandin to progesterone and eCG estrus synchronization treatment.

The objective of this study was to determine the efficacy of PGF2 alpha treatment on pregnancy and calving rates in autumn-calving suckler beef cows synchronized with progesterone and eCG. The population studied consisted of 124 Charolais and 130 Limousin cows in 13 and 12 beef herds, respectively. In each herd, pairs of cows were formed according to parity, body condition score and calving difficulty. Group 1 received a progesterone releasing intravaginal device (PRID) for 12 d with a capsule containing 10 mg estradiol benzoate at implant insertion and 500 IU eCG at PRID removal (Day 0). Group 2 received the same treatment plus 25 mg i.m. dinoprost at Day -2. Each cow was artificially inseminated 56 h after PRID removal (Day 3). Plasma progesterone concentrations were measured to determine cyclicity prior to treatment in samples take on Days -22 and -12, to confirm the occurrence of ovulation (Day 13) and to determine the early pregnancy rate (Day 26). Serum pregnancy-specific protein B (PSPB) concentrations were determined to assess pregnancy rate at Day 39. The effects of variation factors on pregnancy and calving rates after treatment were studied using logistic mixed models and a Cox model, respectively. There were no significant differences between groups or breeds for the rate of cyclicity before treatment nor for ovulation rate (means, 74.1 and 95.7%, respectively). Cyclicity was, however, influenced by individual factors such as body condition score (OR = 3.36, P = 0.001), parity (OR = 5.4, P = 0.001) and herd factors such as stocking rate (OR = 5.62, P = 0.001). The use of a prostaglandin injection increased pregnancy rate at Day 26 (71.7 vs 56.7%, P = 0.01) and at 39 d (67.7 vs 54.3%, P = 0.02) and the calving rate at induced estrus (64.5 vs 48.5%, P = 0.01). We observed 9 twin calvings (5.6%) which occurred in cyclic cows only before treatment. Cows in Group 2 had a 1.5 greater chance of calving before 300 d following the first AI than cows in Group 1 (P = 0.03). In conclusion, the addition of PGF2 alpha injection, 48 h before PRID removal, increased reproductive efficiency in autumn-calving Charolais and Limousin suckler beef cows compared to a classical estrus synchronization treatment using a PRID + eCG.

Influence of flushing on LH secretion, follicular growth and the response to estrus synchronization treatment in suckled beef cows.

The effects of energy supplementation (flushing) on LH and estradiol secretion, follicular growth and the response to estrus synchronization treatment (Norgestomet + PMSG initiated 41.9 +/- 3.4 d after calving) were investigated in 16 suckled beef cows fed either 70% (Group C, n = 8) of energy requirements from calving to 3 wk after AI or fed the same restricted diet until 11 d before synchronization and then were supplemented with 2 kg concentrate until 3 wk after AI (Group S, n = 8). Concentrations of LH and estradiol 17 beta were measured from 3 sampling periods: 25 and 39 d after calving and between 29 and 49 h after implant removal. Ovaries were examined by ultrasonography 11 d before treatment to implant withdrawal (IR). The effects of energy level, day (or hour) of observation and corresponding interactions were tested on repeated measurements by split-plot ANOVA. No positive effect of flushing was observed on characteristics of LH secretion on Day 39. However, the size of the largest follicle and the number of large follicles were higher in Group S than in Group C cows, respectively, 7 and 9 d after the beginning of flushing to 2 d after the start of treatment. After IR, the estradiol secretion tended to be higher in Group S than in Group C cows (9.8 +/- 0.4 pg/mL vs 7.2 +/- 0.2 pg/mL; P = 0.06), but no effect on LH secretion was observed. After implant removal 12 cows ovulated (Group S: 7/8 vs Group C: 5/8; P > 0.05), 7 were pregnant at 21 d after AI (Group S: 6/8 vs Group C: 1/8; P < 0.05) and 4 at 45 d after AI (Group S: 4/8 vs Group C 0/8; P > 0.05). To conclude, flushing had a positive effect on follicular growth, which does not seem to be mediated by LH. In cows fed a restricted diet, flushing enhanced follicular growth, increased the fertilization rate and/or reduced early embryonic death.

Absence of response to oestrus induction and synchronization treatment is related to lipid mobilization in suckled beef cows.

Energy status, follicular growth, oestradiol and LH secretion were investigated in 17 suckled Charolais cows synchronised 59.0 +/- 3.6 days after calving with a 10 day ear implant containing 3 mg of Norgestomet. The cows received 3 mg of Norgestomet and 5 mg of oestradiol valerate by IM injection at implant insertion (day 0) and 600 IU PMSG at implant removal (day 10). They were artificially inseminated (AI) 48 and 72 h after implant removal. Energy status was assessed by measuring weekly plasma concentrations of non-esterified fatty acids (NEFA), beta hydroxy-butyrate (BHB), glucose and insulin 7 weeks before AI. Progesterone plasma concentrations were measured during the same period to assess the presence of a functional corpus luteum. Follicular growth was followed daily by ultrasonography from day -3 to day 13. Oestradiol secretion was measured on day -3, day 6 and day 10 from five hourly samples. Oestradiol and LH plasma concentrations were measured hourly from 29 to 48 h after implant removal for seven cows. Cows were checked for pregnancy by ultrasonography 45 days after AI. Pregnant cows (P) were compared with non-pregnant cows (NP) for energy status, follicular growth, and oestradiol secretion by split-plot ANOVA. Two cows (11.8%) were cyclic before treatment, seven ovulated after treatment (41.2%) and five were found pregnant 45 days after AI (29.4%). There was no difference in body condition score and body weight between P and NP cows on day 0 (2.5 +/- 0.2 and 685 +/- 24 kg vs 2.5 +/- 0.1 and 670 +/- 13 kg; P > 0.05). Mean plasma NEFA concentrations before treatment were significantly lower in P than in NP cows (218 +/- 29 mu eq/L vs 279 +/- 18 mu eq/L; P < 0.05). No significant differences between P and NP cows were found for BHB, glucose and insulin concentrations. P cows presented more medium sized follicles (5 mm < or = diameter < 10 mm) than NP females during the period of observation (2.65 +/- 0.19 vs 2.50 +/- 0.12; P = 0.05). Plasma oestradiol concentrations were not different between P and NP cows on day -3 (8.4 +/- 0.7 pg/mL vs 7.7 +/- 0.4 pg/mL, P > 0.05), day 6 (10.4 +/- 0.6 pg/mL vs 9.8 +/- 0.4 pg/mL, P > 0.05) but were higher in P than in NP cows on day 10 (10.9 +/- 0.6 pg/mL vs 7.8 +/- 0.4 pg/mL; P < 0.05). After implant removal, oestradiol secretion only increased in P cows and a LH peak occurred whereas no increases in oestradiol (11.0 +/- 0.4 pg/mL vs 6.3 +/- 0.3 pg/mL, P < 0.05) and LH (6.0 +/- 0.5 ng/mL vs 1.2 +/- 0.5 ng/mL, P < 0.05) secretion were observed in NP cows. The conclusion was that follicular growth, oestradiol secretion, ovulation and pregnancy rate after oestrus synchronisation treatment are related to mobilization of energy stores before treatment in suckled beef cows in the same body condition score.

Variation factors of pregnancy rates after oestrus synchronization treatment in French Charolais beef cows.

An epidemiological study was conducted in Bourgogne (France) in Charolais herds sampled in 1990 (sample 1:54 herds) and from 1991 to 1993 (sample 2:33 herds). The aim was primarily to quantify, at both herd and individual levels, the factors that result in variations in the response to synchronization treatment as estimated by pregnancy rate after first insemination. The second goal was to test a similar model in sample 2, which included only primiparous cows. In sample 1,329 cows of the 627 studied (52%) became pregnant compared with 122 primiparous cows of the 249 studied (49%) in sample 2. Pregnancy rates after treatment per herd ranged from 0-100% across the herds. Mixed logistic regression was used to calculate the odds ratio (OR) and to take into account both individual and herd level variables for each sample. In the first sample, multiparous cows were more likely to become pregnant after synchronization treatment than primiparous cows (OR = 2.4, P < 0.001). The following individual variables also influenced the response to synchronization: body weight at implant insertion (OR = 1.006, P = 0.017) and calving difficulty (OR = 0.33, P = 0.008). In the second sample, results varied with the year of data collection (OR = 3.46, P = 0.016) and with two herd-level variables: presence of a bull (OR = 0.48, P = 0.047) and type of housing (OR = 2.5, P = 0.004). In conclusion, the results after synchronization depended on both herd-level and individual variables and it was necessary to take into account two levels of factors to reduce variability after treatment.

Influence of postpartum energy restriction on energy status, plasma LH and oestradiol secretion and follicular development in suckled beef cows.

Effects of postpartum energy restriction, parity and time after parturition on energy status (measured by glucose, insulin, non-esterified fatty acids (NEFAs) and beta-hydroxybutyrate), LH secretion and follicular growth were investigated in ten primiparous and nine multiparous suckled cows. Females were allocated by parity, body mass and body condition score at calving to diets supplying either 100% (CE, n = 10) or 70% (LE, n = 9) of energy requirements until day 70 postpartum. Metabolic parameters were measured every week from calving to day 70 postpartum. Blood samples were collected at intervals of 15 min for 10 h on day 30 and day 50 after parturition for LH measurement. Ovaries were examined between days 20 and 30 and days 40 and 50 postpartum by ultrasonography. Energy supply affected mean plasma concentrations of glucose (CE: 0.64 +/- 0.01 g l-1 versus LE: 0.61 +/- 0.01 g l-1; P < 0.05) and NEFA (CE: 168 +/- 17 mu eq l-1 versus LE: 309 +/- 18 mu eq l-1; P < 0.01) but by day 70 postpartum, glucose and NEFA concentrations were not significantly different between the two groups. LH pulse amplitude and frequency were not affected by energy supply (P > 0.10). However, at day 30 postpartum, LH pulse frequency was negatively correlated with plasma concentration of NEFA (r = -0.61; P < 0.01). Cows fed diets supplying 100% of energy requirements had more large follicles than did cows fed low energy diets (CE: 0.82 +/- 0.05 versus LE: 0.31 +/- 0.05; P < 0.05). The size of the largest follicle was greater in CE cows than in LE cows (CE: 10.2 +/- 0.1 mm versus LE: 8.7 +/- 0.2 mm; P < 0.05). Between 40 and 50 days postpartum, the size of the largest follicle was negatively correlated with NEFA concentration (r = -0.5; P < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

Immunological aspects of mammary tumors in dogs and cats: a survey including own studies and pertinent literature.

Naturally occurring cancer in companion animals parallels cancer in man more closely than does experimentally induced cancer in inbred laboratory animals. In dogs and cats, as in man, a role for immune responses is indicated in the development of tumors. A survey is presented based on the literature and our own studies concerning the immunological and immunotherapeutic aspects of canine and feline mammary neoplasia. In dogs bearing mammary neoplasms, circulating immune complexes appear to play a negative role in the generation of effective antitumor immune responses. The functional role of peripheral blood lymphocytes and tumor-infiltrating lymphocytes in dogs and cats with mammary tumors is not yet fully established. No tumor antigen responsible for humoral or cellular responses has yet been identified. Extracorporeal perfusion of serum of dogs with mammary tumors and subcutaneous administration of mitomycin- and neuraminidase-treated autologous tumor cells are associated with improved prognosis. The opposite was true for i.v. treatment with BCG or Corynebacterium parvum vaccine in our study, in contrast to a previous report. A number of other treatment modalities in cats and dogs with mammary carcinomas failed to induce tumor regression. Canine and feline mammary carcinomas are good candidates for modern immunotherapeutic approaches.

Reversible effects of long-term treatment with D-Trp6-LH-RH-microcapsules on pituitary-gonadal axis, spermatogenesis and prostate morphology in adolescent and adult dogs.

The effect of long term treatment with D-Trp6-LH-RH in microcapsules (GnRH-A) on pituitary gonadal axis was studied in a adolescent and adult Fox-terrier dogs. They received intramuscularly 50 micrograms/GnRH-A/kg, on day 1 and 21 and every 4 weeks thereafter. Three adult dogs received 4 injections. cLH, cFSH and T levels were undetectable on day 7. Detectable then normal levels occurred 60 and 90 days respectively after the last injection on day 77. Testis thickness was respectively 22.1 +/- 0.8 mm and 16.3 +/- 0.8 mm on days 0 and 77; initial values were observed 90 days later. Spermatozoa disappeared from the ejaculate on day 21 in 2 dogs; reappearance and complete recovery were observed on days 161-175 and 252 respectively. Histological findings showed on day 91 atrophic lesions of testis and prostate and spermatogonia were present in all seminiferous tubules. After recovery a normal histological appearance was noticed. Three adolescent 29 weeks old dogs received 14 injections, the last one on day 357. cLH, cFSH and T levels were undetectable only from day 105. Testis thickness were respectively 15.8 +/- 0.7 mm, 18.1 +/- 0.7 mm, 12.5 +/- 0.3 mm and 21.4 +/- 0.7 mm on day 0, 21, 357 and 490. Initially, no spermatozoa were present in the ejaculates, they appeared in 2 dogs after 2 months for 20 to 40 days then disappeared until day 449. Normal semen characteristics were observed in all three dogs on day 581. Histological findings on day 371 were comparable to those observed in the adult dogs. This study demonstrates that longterm treatment with D-Trp6-LH-RH in microcapsules leads to a reversible inhibition of spermatogenesis in dogs. The delayed response in adolescent dogs might be due to a transient resistance to therapy related to.

Plasma concentration of luteinizing hormone, testosterone, dehydroepiandrosterone, androstenedione between birth and one year in the male dog: longitudinal study and hCG stimulation.

A longitudinal study from peripheral blood, with samples collected every week, was performed between birth and one year of age on young Fox terriers dogs in order to determine the patterns of plasma LH, T, DHA and A concentrations. T, DHA, A curves show the same profile. The dog model shows the 3 successive steps preceding the adult life that are met in one year time: First, an infantile period between birth and the 12th week of age: the basal level of LH (4.29 ng/ml) and the 3 androgens levels (T less than 0.3 ng/ml, DHA less than 0.45 ng/ml, A less than 0.36 ng/ml) are low. A pubertal period, between the 13th week and the 36th week of age: we observe the maximum activity of the pituitary gland. The basal level of LH (7.97 ng/ml) significantly increase (P less than 0.001). The mean levels of androgens from 17 to 27 weeks of age are still quite low although significantly higher (P less than 0.03) for T and DHA than previously. After the 27th week of age, the androgens concentrations drastically increase. A post pubertal period begins at the 36th week of age. The mean LH (5.85 nh/ml) decrease. The androgens concentrations seem to plateau during the 12th month of age in the range of 2.5-5 ng/ml for T, 1.5-2.5 ng/ml for DHA and 1-2 ng/ml for A. hCG test (35 UI/kg, IM), with samples collected at 6, 12, 24, 30, 36 and 48 hours post injection, were performed at 1, 4, 7, 9 and 12 months of age. At 1 month, only minor variations were noticed; but after 4 months of age, for the 3 steroids, the same time course response was observed as in the adult dog: maximum levels were reached earlier for DHA (6-12 h) than for T and A (24 h).

Growth, testicular development and sperm output in the dog from birth to post pubertal period.

This study was performed in Fox terrier dogs. Their mean body weight at birth is 315 g; it reaches 12 kg at 1 year of age. The body growth pattern has a sigmoid appearance with a rapid growth phase between 9 weeks and 24 weeks of age. With frequent radiographic examinations, 39 ossification centers were individualized on the long bones; they had all appeared by 25 weeks of age. The fusion of these centers is achieved between 36 and 44 weeks of age. This determination of the bone age has not proved to be a good factor to date the morphologic events of puberty. The testicular width measure is a reliable criteria which is well correlated to the other morphologic and spermatic criteria (volume of ejaculate, spermatozoa concentration). The testicular growth is rapid (width: 0,6 mm/week) between 22 and 36 weeks of age. The first spermatozoa are observed at the end of this period (32 to 34 weeks of age). During the post pubertal period, from 34 weeks to 45 weeks of age, volume of ejaculate, motility, spermatozoa concentration increase and percentage of dead spermatozoa and abnormal forms decrease.

Animal models for hormone-dependent human breast cancer. Relationship between steroid receptor profiles in canine and feline mammary tumors and survival rate.

The present study shows that canine and feline mammary tumors, like human breast tumors, can be polyreceptive, i.e., they can contain estrogen (ER), progestin (PR), androgen, glucocorticoid, and/or mineralocorticoid cytosol receptors. Furthermore, a follow-up of 45 bitches with mammary carcinoma has indicated that the survival rate is significantly higher in animals with receptor-rich (ER and/or PR) tumors. This indicates that these canine mammary tumors should be evaluated further for their suitability as an animal model for hormone-dependent human breast carcinoma.

Intratumoral BCG and Corynebacterium parvum therapy of canine mammary tumours before radical mastectomy.

In two parallel studies, bitches with mammary tumour received single intralesional injections of BCG (1 mg: 10(7) living bacteria) and Corybacterium parvum (10(9) killed bacteria) (53 bitches) or C. parvum alone (129 bitches) at the same dosage. Control groups received injections, following the same protocol, of 1 ml BCG suspension medium diluted in saline in the first study (51 bitches) or no injections at all (120 bitches in the second study). A block dissection, including mammary tumours, adjacent mammary glands, and regional lymph nodes, was performed 2 weeks later in all animals. On the basis of histologically confirmed malignant tumours, 48 bitches (25 treated by-immunotherapy and 23 controls) in the first study and 67 bitches (30 treated by immunotherapy and 37 controls) in the second study remained for postsurgical follow-up. The clinical tolerance of the treatment was generally good. No significant differences were found in cumulative survival rates between treated and control group in either studies.